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- Differential Sensitivity of Taxol-induced Apoptosis in U2OS and SaOS2 Osteogenic Sarcoma Cells
- Jung Hye Kim, Byung Rho Chin, Seong Yong Kim, Jae Ryong Kim, Suk Hwan Baek
- Cancer Res Treat. 2003;35(2):148-153. Published online April 30, 2003
- DOI: https://doi.org/10.4143/crt.2003.35.2.148
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Abstract
PDF - PURPOSE
Taxol (Paclitaxel) is a new generation of chemotherapeutic drug proven to be effective in the treatment of many cancers. In this study, to further demonstrate the differential effect of the tumor suppressor gene, p53, on the Taxol-induced apoptosis in osteogenic sarcoma cell lines, we used p53-defected SaOS2 cells and wild type p53-expressed U2OS cells. MATERIALS AND METHODS: The cell viability was measured by the XTT assay. To examine whether the differential expressions of p53, in U2OS and SaOS2 cells, were associated with Taxol-induced apoptosis, DNA fragmentation assays were performed on both cytosolic and genomic DNA. Since the cleavage of poly (ADP-ribose) polymerase (PARP) is primarily responsible for apoptosis, the cleavage of PARP, and the expression of cyclin B1, polo-like kinase, Bax, Bcl-xL, Bcl-2 in U2OS and SaOS2 cells were compared by Western blot analyses. RESULTS: The cell viability of the p53-defected SaOS2 cells was markedly decreased with Taxol treatment. Whereas, the cell viabilities due to 6-mercaptopurine and adriamycin were no different between the U2OS and SaOS2 cells. Treatment with Taxol induced a ladder- like pattern of DNA fragments, which is a biochemical hallmark of apoptosis, consisting of multiples of approximately 180-200 base pairs, in a dose-dependent manner in the SaOS2 cells, but insignificantly with the U2OS cells. When the cells were treated with Taxol, the 89 kDa cleavage product of PARP clearly appeared as a function of time in the SaOS2 cells, but not in the U2OS cells. The Taxol-induced apoptosis in p53 defected-osteogenic sarcoma cells was associated with the PARP cleavage as a result of the increased activity of caspase 3, and the high expressions of cyclin B1 and PLK. Bax, as a proapoptotic factor, was increased in the SaOS2cells, but the Bcl-xL and Bcl-2 were decreased when the cells were exposed to 10miceoM Taxol. CONCLUSION: From these results, it was concluded that p53-defected SaOS2 cells are much more sensitive to Taxol-induced apoptosis than p53-expressed U2OS cells. -
Citations
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- Imatinib-Functionalized Galactose Hydrogels Loaded with Nanohydroxyapatite as a Drug Delivery System for Osteosarcoma: In Vitro Studies
Paulina Sobierajska, Benita Wiatrak, Paulina Jawien, Maciej Janeczek, Katarzyna Wiglusz, Adam Szeląg, Rafal J. Wiglusz
ACS Omega.2023; 8(20): 17891. CrossRef - Programmed cell death, redox imbalance, and cancer therapeutics
Xiaofeng Dai, Danjun Wang, Jianying Zhang
Apoptosis.2021; 26(7-8): 385. CrossRef - High-Content, High-Throughput Screening for the Identification of Cytotoxic Compounds Based on Cell Morphology and Cell Proliferation Markers
Heather L. Martin, Matthew Adams, Julie Higgins, Jacquelyn Bond, Ewan E. Morrison, Sandra M. Bell, Stuart Warriner, Adam Nelson, Darren C. Tomlinson, Maria A. Deli
PLoS ONE.2014; 9(2): e88338. CrossRef - The Time‐Dependent Serial Gene Response to Zeocin Treatment Involves Caspase‐Dependent Apoptosis in HeLa Cells
Jooyeon Hwang, Young‐Youl Kim, Sungjin Huh, Junghee Shim, Chan Park, Kuchan Kimm, Dong Kug Choi, Tae‐Kyu Park, Soonhag Kim
Microbiology and Immunology.2005; 49(4): 331. CrossRef - Proteome Analysis of Differential Protein Expression in Cervical Cancer Cells after Paclitaxel Treatment
Eun-Kyoung Yim, Jun-Sang Bae, Seung-Bak Lee, Keun-Ho Lee, Chan-Joo Kim, Sung-Eun Namkoong, Soo-Jong Um, Jong-Sup Park
Cancer Research and Treatment.2004; 36(6): 395. CrossRef
- Imatinib-Functionalized Galactose Hydrogels Loaded with Nanohydroxyapatite as a Drug Delivery System for Osteosarcoma: In Vitro Studies
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- Role of ATF on Transcriptional Regulation of DNA Topoisomerase II a Gene in HL - 60 Arrested to G2 / M and M Phase
- Kyu Lim, Mee Young Son, Byung Ik Choi, Kyung Ah Yun, Meizi Zheng, Tae Wook Kang, Young Chul Lee, Jong II Park, Wan Hee Yoon, Byung Doo Hwang
- J Korean Cancer Assoc. 1999;31(6):1279-1287.
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Abstract
PDF
- PURPOSE
To gain insight on transcriptional repression of Topo II a in HL-60 cells arrested to G2/M and M phase, the levels of Topo IIa mRNA and the binding activity of ATF have been investigated with Northern blot hybridization and DNA mobility shift assay, respectively.
MATERIALS AND METHODS
HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-mactivated fetal bovine serum and antibiotics in a humidified 5% CO2 at 37C degree. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. A Xho I-Mlu I fragment of phTOP2 was used as probe for Northern blot analysis of Topo II a mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA oligomer (upper strand, 5-TCTCCGCTATGACGCCGAGTGGTG-3) for ATF binding activity was mixed with nuclear extracts in a 20 pl reaction volume containing 60 mM KC1, 12 mM HEPES, pH 7.9, 5 mM MgCl2, 0.2 mM EDTA, 0.2 mM DTT, 12% glycerol, and 2 ug of poly [dI-dC].
RESULTS
HL-60 cells were arrested at G2/M phase and M phase after taxol or nocodazole treatment. The levels of Topo II a mRNA were reduced at 24 hours after exposure with nocodazole or taxol but the unknotting activities were not changed. DNA mobility shift assay using oligonucleotide containing the ATF binding site showed that ATF binding activity was reduced after pretreatment of nododazole or taxol.
CONCLUSIONS
These results suggest that the reduction of ATF binding activity may be important to transcriptional repression of Topo II a gene by nocodazole and taxol in HL- 60 cells.
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